U.S. patent application number 11/995958 was filed with the patent office on 2010-05-27 for method to inhibit proliferation and growth of metastases.
This patent application is currently assigned to TRUSTEES OF BOSTON UNIVERSITY. Invention is credited to Michael T Kirber.
Application Number | 20100130904 11/995958 |
Document ID | / |
Family ID | 37451525 |
Filed Date | 2010-05-27 |
United States Patent
Application |
20100130904 |
Kind Code |
A1 |
Kirber; Michael T |
May 27, 2010 |
METHOD TO INHIBIT PROLIFERATION AND GROWTH OF METASTASES
Abstract
The present invention provides a method to reduce the amount of
undesired growth factors in the circulating blood of a subject to
prevent tumor growth and proliferation during or after a wound
healing and/or other local tissue repair process on a subject,
comprising extracorporeal adsorption of growth factors from blood
of the subject and return of the treated blood to the subject.
Inventors: |
Kirber; Michael T; (Sharon,
MA) |
Correspondence
Address: |
RONALD I. EISENSTEIN
100 SUMMER STREET, NIXON PEABODY LLP
BOSTON
MA
02110
US
|
Assignee: |
TRUSTEES OF BOSTON
UNIVERSITY
Boston
MA
|
Family ID: |
37451525 |
Appl. No.: |
11/995958 |
Filed: |
July 18, 2006 |
PCT Filed: |
July 18, 2006 |
PCT NO: |
PCT/US06/27746 |
371 Date: |
September 11, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60700118 |
Jul 18, 2005 |
|
|
|
Current U.S.
Class: |
604/5.01 |
Current CPC
Class: |
A61M 1/3486 20140204;
A61M 1/3472 20130101; A61M 1/3679 20130101; C07K 16/22
20130101 |
Class at
Publication: |
604/5.01 |
International
Class: |
A61M 1/36 20060101
A61M001/36 |
Claims
1. A method for reducing the amount of at least one circulating
growth factor from circulating blood of a subject with a tissue
injury, comprising contacting at least a portion of the circulating
blood of a subject affected with a tissue injury with an
extracorporeal adsorption device wherein the device comprises at
least one adsorption compound that binds to at least one growth
factor in the circulating blood of the subject.
2. The method of claim 1, wherein the tissue injury is caused by
surgery.
3. The method of claim 2, wherein the surgical wound is induced
during removal of a tumor.
4. The method of claim 1, wherein the tissue injury is caused by
radiation therapy.
5. The method of claim 1, wherein the subject has or is at risk for
developing cancer.
6. The method of claim 1, wherein at least one growth factor is
selected from the group consisting of TGF-.alpha., TGF-.beta.1,
TGF-.beta.2, TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2, FGF-1, FGF-2
basic, VEGF, TNF-.alpha., FGF-7 and any combination thereof.
7. A method of treating a subject undergoing wound healing,
comprising contacting the blood of said subject with an
extracorporeal adsorption device wherein the device contains
adsorption compounds that bind to growth factors in the subject's
blood.
8. The method of claim 7, wherein the subject is affected with a
surgical wound.
9. The method of claim 7, wherein the subject is affected with a
surgical wound induced during surgical removal of a tumor.
10. The method of claim 7, wherein the subject is at risk for
cancer.
11. The method of claim 7, wherein the growth factors are selected
from the group consisting of FGF-1/FGF acidic, FGF-2/FGF basic,
FGF-3, FGF-4, FGF-5, FGF-6, FGF-7/KGF, FGF-8, FGF-9, FGF-10,
FGF-11, FGF-12, FGF-12, FGF-13, FGF-14, FGF-15, FGF-16, FGF-17,
FGF-18, FGF-19, FGF-20, FGF-21, FGF-22, FGF-23, IL-1.alpha.,
IL-1.beta., IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10,
IL-11, IL-12; G-CSF, M-CSF/CSF-1, GM-CSF, EGF, HB-EGF,
Amphiregulin, Betacellulin, Epigen, Epiregulin, NRG-3, NRG1 isoform
GGF2, NRG1 Isoform SMDF, NRG1-alpha/HRG1-alpha, NRG1-beta
1/HRG1-beta 1, TMEFF1, TMEFF2, TGF-.alpha., VEGF, VEGF-B, VEGF-C,
VEGF-D, PIGF-1, PIGF-2, PIGF-3, PDGF, PDGF-A, PDGF-B, PDGF-C,
PDGF-AB, Neuropilin-1, Neuropilin-2, TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, TGF-.beta.4, TGF-.beta.5, Schwann cell-derived Growth
Factor, NGF, IGF-1 and IGF-2, Glial Growth Factor, TNF-.alpha.,
TNF-.beta., CTGF/CCN2, NOV/CCN3, PD-ECGF/gliostatin, EG-VEGF/PK1,
Hepassocin, HGF/hepapoietin A/scatter factor, .beta.-NGF,
Progranulin, Thrombopoietin, Prolactin, Prostaglandins, GH1, GH2,
and any combination thereof.
12. The method of claim 7, wherein the growth factors are selected
from the group consisting of TGF-a, TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2, FGF-1/FGF acidic, FGF-2/FGF
basic, VEGF, TNF-.alpha., FGF-7/KGF, and any combination
thereof.
13. A method of treating an individual undergoing removal of a
cancer comprising removing undesired circulating growth factors
from the individual.
14. The method of claim 13, wherein the circulating growth factors
are removed by an extracorporeal adsorption device.
15. The method of claim 14, wherein the device comprises at least
one adsorption compound that binds to at least one growth factor in
the circulating blood of the individual.
16. The method of claim 13, 14 or 15, wherein the undesired
circulating growth factor is selected from the group consisting of
TGF-.alpha., TGF-.beta.1, TGF-.beta.2, TGF-.beta.3, PDGF, EGF,
IGF-1, IGF-2, FGF-1, FGF-2, VEGF, TNF-.alpha., FGF-7 and any
combination thereof.
17. The use of an extracorporeal device to remove at least one
undesired circulating growth factor from the blood of an individual
with a tissue injury.
18. The use of claim 17, wherein the tissue injury is caused by
surgery or radiation therapy.
19. The use of claim 17 or 18, wherein the subject is at risk for
cancer.
20. The use of claim 19, wherein the subject had a cancer.
21. The use of claim 17, 18, 19, or 20 wherein the undesired
circulating growth factor is selected from the group consisting of
TGF-.alpha., TGF-.beta.1, TGF-.beta.2, TGF-.beta.3, PDGF, EGF,
IGF-1, IGF-2, FGF-1, FGF-2, VEGFF, TNF-.alpha., FGF-7 and any
combination thereof.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims the benefit under 35 U.S.C.
.sctn.119(e) of U.S. provisional application No. 60/700,118, filed
Jul. 18, 2005, which application is herewith incorporated by
reference in its entirety.
BACKGROUND OF THE INVENTION
[0002] It is well known that surgical removal of a primary tumor is
frequently followed by the rapid growth of multiple metastases and
often death. When a subject is evaluated and metastases are found,
removal of the primary lesion is often no longer considered the
best course of treatment. Proliferation of metastases following
removal of the primary lesion has led to the hypothesis that the
primary lesion releases or causes the release of anti-angiogenic
molecules, which block the growth of metastases. This has been an
area of active research resulting in the identification of
substances which may be useful in blocking tumor growth (for
example, endostatin, angiostatin).
[0003] New forms of cancer treatment have involved blocking the
effects of growth factors such as vascular endothelial growth
factor (VEGF), fibroblast growth factor (FGF), platelet derived
growth factor (PDGF), epidermal cell growth factor (EGF) and
others. These agents can be receptor blockers such as monoclonal
antibodies to the receptors (for example, HERCEPTIN.TM.
(Trastuzumab), IMC-1121b), other competitive receptor-binding
agents (for example, ERBITUX.TM. (cetuximab)). The agents can also
block the activity of a growth factor receptor by interfering with
its ability to function without necessarily blocking the
extracellular binding site. For example, TARCEVA.RTM. (erlotinib)
and IRESSA.RTM. (gefitinib) are thought to block the intracellular
tyrosine kinase domain of the EGF receptor.
[0004] Following tissue injury caused by, for example, surgery or
radiation therapy, circulating levels of the growth factors can
become and are frequently elevated. This period of growth factor
elevation during wound healing and/or inflammation may be a key
factor in triggering the proliferation of metastases and
micrometastases following surgery such as a surgery for treatment
of cancer. However, blocking growth factors following surgery or
radiation therapy could interfere with wound healing and
consequently would not be a desirable course of treatment.
[0005] Thus, there is a need for methods to reduce systemic
increase and circulation of growth factors in the body following
surgery or other tissue injury without interfering with local
surgical wound healing and/or repair of local tissue injury, such
as injury caused by radiation therapy.
SUMMARY OF THE INVENTION
[0006] The present invention provides a method to reduce the amount
of undesired growth factors in the circulating blood of a subject
to prevent tumor growth and proliferation during or after a wound
healing and/or other local tissue repair process on a subject,
comprising extracorporeal adsorption of growth factors from blood
of the subject and return of the treated blood to the subject.
[0007] Any known means for extracorporeal removal can be used. For
example, an apparatus for extracorporeal circulation of whole blood
or plasma is connected to a subject. Growth factors can be removed
from the blood or plasma in the apparatus by using a means for
removing the growth factors such as affinity adsorption. One can
use antibodies (including fragments thereof), growth factor
receptors, nucleic acids, small molecules, molecules containing
receptor or antibody mimetics, and the like to bind to the desired
growth factor and remove that factor from the circulating blood or
plasma. This systemic removal will not significantly affect the
local levels of growth factors near a wound.
[0008] In one embodiment, the method is performed on a subject who
is undergoing surgical removal of a tumor prior to administration
of the methods of the present invention.
[0009] In one embodiment, the method is performed on a subject who
underwent surgery unrelated to cancer prior to administration of
the methods of the present invention.
[0010] In one embodiment, the method is performed on a subject who
is at risk for cancer.
[0011] In one embodiment, the method is performed on a subject who
is at risk for metastasis.
[0012] The present invention provides a method to prevent and/or
inhibit metastatic and micrometastatic tumor growth in a subject
undergoing surgical wound healing (i.e. treat a subject undergoing
wound healing), comprising elimination of systemically circulating
growth factors via extracorporeal adsorption of growth factors and
return of blood to the subject.
[0013] In one embodiment, the surgical wound is a result of removal
of a tumor.
DETAILED DESCRIPTION OF THE INVENTION
[0014] The present invention is based on the discovery that the
amount of growth factors can be reduced in systemic circulation
without significantly altering local levels of growth factors near
a site of a tissue injury, thereby preventing metastatic tumor
growths following tissue injury, for example, surgery and radiation
therapy.
[0015] The present invention provides a method to treat a subject
by reducing the amount of undesired growth factors from the
circulating blood of a subject, comprising extracorporeal
adsorption of growth factors and return of blood to the
subject.
[0016] In one embodiment, the method is performed in conjunction
with a surgical removal of a tumor. In one embodiment, the method
may be performed prior to surgery, during surgery, or after
surgery.
[0017] In one embodiment at least 1, 2, 3, 4, 5 or more removal
treatments are performed in conjunction with the surgical
proceeding. One can begin this procedure at any time prior to
surgery, for example, 24 hours, 8 hours, 5 hours, 4 hours, 3 hours,
2 hours, 1 hours, 0.5 hours or less. One can perform the procedure
one or more times, or continuously during the surgery. One can also
continue this procedure indefinitely. Typically, one performs one
or more rounds of removal during a period of at least 8 hours, 24
hours, 36 hours, 48 hours, 72 hours and up to one week, 2 weeks, 3
weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or even up to 2 months, 3
months, 4 months, 5 months or 6 months. One can begin the procedure
following the surgery. Preferably one begins the treatment within
at least one hour to one week of the surgery, for example within
the first two days, preferably within the first day. All time
periods in between are part of the procedure. In one preferred
embodiment, treatment is completed during a 72 hour period from
surgery. One does not have to use this method continually, but
rather periodically to prevent build up of undesired circulating
growth factors. Levels of growth factors can be determined by known
means.
[0018] In one embodiment, the method is performed in conjunction
with a surgery unrelated to cancer.
[0019] In one embodiment, the method is performed in conjunction
with radiation therapy.
[0020] In one embodiment, the subject is at risk for cancer.
[0021] In one embodiment, the subject is at risk for
metastasis.
[0022] The present invention provides a method to inhibit/prevent
tumor growth, particularly metastasis and micrometastasis, in a
subject undergoing surgical wound healing comprising, reduction or
elimination of systemically circulating growth factors via
extracorporeal adsorption of growth factors from blood and return
of blood to the subject. In one embodiment, the surgical wound is a
result of removal of a tumor.
[0023] In the methods of the present invention, one reduces the
amount of circulating growth factors from the blood system using
extracorporeal means. This is done by standard means, for example,
an apparatus for extracorporeal circulation of whole blood or
plasma is connected to the subject through tubing lines and blood
access device(s). Such an apparatus should provide conduits for
transporting the blood to an adsorption device and conduits for
returning the processed blood or plasma to the subject. When plasma
is processed through the adsorption device, a plasma separation
device is typically used as well as means of mixing the
concentrated blood with processed plasma. The later is normally
achieved by leading the two components into an air-trap where the
mixing occurs.
[0024] Devices for extracorporeal affinity adsorption have been
developed by Aethlon Medical Inc. (San Diego, Calif.) for the
removal of materials such as HIV and other viruses from blood (See
U.S. Pat. No. 6,528,057 and U.S. Pat. App. No. 2204/0175291, which
are incorporated herein in their entirety). Mitra Medical AB (Lund,
Sweden) has developed an extracorporeal affinity adsorption device
to filter labeled antibodies from the blood (See U.S. Pat. Nos.
6,723,318, 6,558,543, 6,251,394, 4,965,112, E.P. 0436717, Int'l
Pat. App. Nos. WO 05/4615, WO 05/051424, WO 04/022111, WO 01/95857,
and U.S. Pat. App. Nos. 2004/0052784, 2002/0159994, 2001/0023288,
which are herein incorporated in their entirety). Other
extracorporeal affinity adsorption devices have been disclosed in,
for example, U.S. Pat. Nos. 4,714,556 and 4,787,974 to Ambrus and
Csaba; 6,099,730 to Ameer et al.; and U.S. Pat. Nos. 6,039,946,
6,569,112, 6,676,622, and U.S. Pat. App. No. 2004/0220508 to
Strahilevitz. Grovender et al. (Kidney International. (2004)
65:310-322) describe extracorporeal removal of
.beta..sub.2-microglobin using a single-chain antibody
fragment-based immunoadsorbant device. The methods of the present
invention contemplate use of the extracorporal affinity adsorption
devices disclosed in the above publications and others known to the
skilled artisan.
[0025] In one embodiment, one uses the method to reduce the amount
of circulating growth factors other than cytokines. In one
embodiment, one does not reduce the amount of TNF-.alpha.,
IL-1.beta. or IL-6. Extracorporeal techniques for blood clearance
are widely used in kidney dialysis, where toxic materials build up
in the blood due to the lack of kidney function. Other medical
applications, in which an extracorporeal apparatus can be used,
include: removal of radioactive materials; removal of toxic levels
of metals, removal of toxins produced from bacteria or viruses;
removal of toxic levels of drugs, and removal of whole cells (for
example, cancerous cells, specific haematopoietic cells such as B,
T, or NK cells) or removal of bacteria and viruses.
[0026] One can adapt the extracorporeal affinity adsorption device
for the adsorption of the desired growth factor based upon the
present specification. For example, the desired growth factors may
be adsorbed through the use of antibodies, such as Fab', monoclonal
antibodies, single chain antibodies, antibody fragments, etc.,
nucleic acids, small molecules, growth factor receptors, molecules
containing receptor or antibody mimetics, any selective high
affinity molecule or combinations thereof. The requirement is that
such affinity compounds bind the growth factors that one wishes to
reduce or remove from the circulation. Multiple species of affinity
compounds may be used simultaneously in the methods of the
invention. Single species of affinity compounds or multiple sets of
species of affinity compounds may bind different growth
factors.
[0027] The extracorporeal affinity device may contain a matrix (as
in the device developed by Mitra Medical AB (Lund, Sweden)) that
may be coated with a ligand, for example, an affinity compound or
affinity compounds that bind growth factors. The extracorporeal
affinity device may contain a compartment wherein the affinity
compounds are contained (see, for example, U.S. Pat. No.
6,099,730). Here, the affinity compounds may be contained as
immobilized on agarose, polyacrylamide or other composition of
beads, or on micelles. The device may also comprise an
ultrafiltration membrane wherein affinity compounds may be
immobilized (see, for example, Aethlon Medical Inc. (San Diego,
Calif.)). Other extracorporeal affinity adsorption devices also
contain similar components on which affinity compounds (for
example, bound to solid support) may be immobilized.
[0028] There is considerable evidence indicating that the overall
process of tissue injury related to inflammation, healing and
repair of damaged tissue (for example, wounds resulting from
surgery or radiation therapy), including the necessary
intercellular communication, is regulated in a coordinated manner
in adult humans and other mammals by a number of specific soluble
growth factors which are released within the wound environment.
Such factors are released, for example, by degranulating platelets
and incoming macrophages. These growth factors contribute to
inducing neovascularisation, leucocyte chemotaxis, fibroblast
proliferation, migration and deposition of collagen and other
extracellular matrix molecules within the site of the injury.
Growth factors that have been identified and isolated are generally
specialized soluble proteins or polypeptides and include, but are
not limited to, transforming growth factor alpha (TGF-.alpha.),
transforming growth factor beta (TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3 etc), platelet derived growth factor (PDGF), epidermal
growth factor (EGF), insulin-like growth factors I and II (IGFI and
IGFII) and acidic and basic fibroblast growth factors (acidic FGF
and basic FGF). General reviews on growth factors can be found, for
example, in articles by Mary H McGrath in Clinics in Plastic
Surgery, Vol. 17, No. 3, July 1990, pp 421-432, and by George A
Ksander in Annual Resorts in Medicinal Chemistry, 1989, Chap, 24
(published by Academic Press, Inc.) of which the contents are
incorporated herein by reference. Growth factor activity is crucial
for proper healing of injured tissue. However, increased systemic
growth factor concentration in blood may result in harmful cell
proliferation.
[0029] For example, many new cancers are initiated, and existing
cancers and hyperproliferative disorders stimulated, by growth
factors that affect either the cancer cell itself, or normal tissue
around the cancer that facilitate survival of the cancer cell (for
example, angiogenic factors). There is a direct correlation between
the circulating level of certain growth factors and cancer
proliferation. Stimulation by high levels of circulating growth
factors released during a body's natural response to a tissue
injury, such as surgical wound healing or radiation therapy, may
increase a subject's risk for tumor growth and metastasis.
[0030] The present invention provides a method of preventing
metastases by reducing the level of circulating growth factors in a
subject to inhibit or prevent tumor growth and cancer recurrence,
and to reduce or eliminate existing cancers, and metastatic and
micrometastatic cell proliferation. Reduction of systemically
circulating growth factors provides a novel way to prevent
metastatic growth of secondary tumors following tissue injury
caused by surgery or radiation therapy. Similarly, a subject who is
at risk for cancer or at risk for recurrence of a cancer will
benefit from the elimination of systemically circulating growth
factors subsequent to surgery.
[0031] Reduction or removal of circulating growth factors by
extracorporeal adsorption reduces systemic levels of circulating
growth factors without significantly altering growth factor levels
at the local site of tissue injury.
[0032] "Metastasis" and "micrometastis" refer to a focus of
cancerous cells related to a preexisting cancer, referred to as
primary tumor or cancer, but that developed remotely from this
primary focus without continuity with it. The dissemination of
these secondary foci typically takes place via lymphatic or hematic
routes.
[0033] As used herein, the term "antibody" (Ab) refers to mammalian
monoclonal antibodies, polyclonal antibodies, multispecific
antibodies (for example, bispecific antibodies), antibody
fragments, immunoglobulin chains or fragments thereof, such as Fv,
Fab, Fab', F(ab').sub.2 or other antigen-binding sub-sequences of
anti bodies, "single-chain Fv" antibody fragments or "diabodies",
so long as they exhibit the desired biological activity, for
example, growth factor binding.
[0034] The term "monoclonal antibody" as used herein refers to an
antibody obtained from a population of substantially homogeneous
antibodies, i.e., the individual antibodies comprising the
population are identical except for possible naturally occurring
mutations that may be present in minor amounts. Monoclonal
antibodies are highly specific, being directed against a single
antigenic site.
[0035] Furthermore, in contrast to conventional (i.e. polyclonal)
antibody preparations which typically include different antibodies
directed against different determinants (epitopes), each monoclonal
antibody is advantageous in that they are synthesized by the
hybridoma culture, uncontaminated by other immunoglobulins. The
modifier "monoclonal" indicates the character of the antibody as
being obtained from a substantially homogeneous population of
antibodies, and is not to be construed as requiring production of
the antibody by any particular method. For example, the monoclonal
antibodies to be used in accordance with the present invention may
be made by the hybridoma method first described by Kohler et al.,
Nature, 256:495 (1975), or made by recombinant DNA methods (see,
for example, U.S. Pat. No. 4,816,567). The monoclonal antibodies
may also be isolated from phage antibody libraries using the
techniques described in, for example, Clackson et al., Nature,
352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597
(1991).
[0036] The monoclonal antibodies herein specifically include
"chimeric" antibodies (immunoglobulins) in which a portion of the
heavy and/or light chain is identical with or homologous to
corresponding sequences in antibodies derived from a particular
species or belonging to a particular antibody class or subclass,
while the reminder of the chain(s) is identical with or homologous
to corresponding sequences in antibodies derived from another
species or belonging to another antibody class or subclass, as well
as fragments of such antibodies, so long as they exhibit the
desired biological activity (U.S. Pat. No. 4,816,567; Morrison et
al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).
[0037] "Antibody fragments" comprise a portion of an intact
antibody, generally the antigen-binding or variable region of the
intact antibody. Examples of antibody fragments include Fab, Fab',
F(ab')2, and Fv fragments, diabodies, single-chain antibody
molecules, and multi-specific antibodies formed from antibody
fragments.
[0038] "Single-chain Fv" antibody fragments comprise the VH and VL
domains of antibody, wherein these domains are present in a single
polypeptide chain. Generally, the Fv polypeptide further comprises
a polypeptide linker: between the VH and VL domains which enables
the sFv to form the desired structure for antigen binding. For a
review of sFv, see Pluckthun in The Pharmacology of Monoclonal
Antibodies, vol. 113, Rosenbourg and Moore eds., Springer-Verlag,
New York, pp. 269-315 (1994).
[0039] The term "diabodies" refers to small antibody fragments with
two antigen-binding sites, which fragments comprise a heavy-chain
variable domain (VH) connected to a light-chain variable domain
(VL) in the same polypeptide chain (VH-VL). By using a linker that
is too short to allow paring between the two domains on the same
chain, the domains are forced to pair with the complementary
domains of another chain and create two antigen binding sites.
Diabodies are described more fully in, for example, U.S. Pat. No.
5,591,828; WO 93/11161; and Hollinger et al., Proc. Natl. Acad.
Sci. USA, 90:6444-6448 (1993).
[0040] As used herein, the term "affinity compound" includes any
composition which binds specifically to a growth factor of the
methods of the present invention. A binding composition or agent
refers to a molecule that binds with specificity to the growth
factor, for example, in a ligand-receptor type fashion or an
antibody-antigen interaction, for example, proteins which
specifically associate with the growth factor, for example, in a
natural physiologically relevant protein-protein interaction,
either covalent or non-covalent. The term "binding composition"
includes small organic molecules, nucleic acids and polypeptides,
such as a full antibody (preferably an isolated monoclonal human
antibody) or antigen-binding fragment thereof. Antibodies and
antigen binding fragments thereof, include, but are not limited to,
monoclonal antibodies, polyclonal antibodies, bispecific
antibodies, Fab antibody fragments, F(ab)2 antibody fragments, Fv
antibody fragments (for example, VH or VL), single chain Fv
antibody fragments and dsFv antibody fragments. In one embodiment,
antibodies may be fully human antibodies or chimeric antibodies.
Preferably, the antibody molecules are isolated monoclonal, fully
human antibodies.
[0041] "Treatment" refers to both therapeutic treatment and
prophylactic or preventative measures. Those in need of treatment
include those already with the disorder as well as those in which
the disorder is to be prevented.
[0042] "Subject" generally refers to a mammal. For purposes of
treatment, mammal refers to any animal classified as a mammal,
including humans, domestic and farm animals, and zoo, sports, or
pet animals, such as dogs, horses, cats, cows, etc. Preferably, the
mammal is human.
[0043] The terms "cancer" and "cancerous" refer to or describe the
physiological condition in mammals that is typically characterized
by unregulated cell growth and is intended to refer to both
malignant and benign extreme or unregulated cell growth. Examples
of cancer include, but are not limited to, carcinoma, lymphoma,
blastoma, sarcoma, and leukemia. More particular examples of such
cancers include squamous cell cancer, small-cell lung cancer,
non-small cell lung cancer, gastrointestinal cancer, pancreatic
cancer, glioblastoma, cervical cancer, ovarian cancer, liver
cancer, bladder cancer, hepatoma, breast cancer, colon cancer,
colorectal cancer, endometrial carcinoma, salivary gland carcinoma,
kidney cancer, renal cancer, prostate cancer, vulval cancer,
thyroid cancer, hepatic carcinoma and various types of head and
neck cancer. The term "tumor" is used interchangeably with "cancer"
herein.
[0044] "Growth factors" the removal/reduction of which is useful
according to the methods of the present invention include growth
factors, and polypeptide angiogenesis factors; and naturally
modified derivatives and naturally occurring peptide fragments
thereof. Growth factors induce or promote cell proliferation and/or
angiogenesis. Growth factors contemplated by the invention
comprise: Fibroblast Growth Factor (FGF) family members including,
but not limited to, FGF-1/FGF acidic, FGF-2/FGF basic, FGF-3,
FGF-4, FGF-5, FGF-6, FGF-7/KGF, FGF-8, FGF-9, FGF-10, FGF-11,
FGF-12, FGF-12, FGF-13, FGF-14, FGF-15, FGF-16, FGF-17, FGF-18,
FGF-19, FGF-20, FGF-21, FGF-22, FGF-23; Interleukins, including but
not limited to, IL-1.alpha., IL-1.beta., IL-2, IL-3, IL-4, IL-5,
IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, and IL-12; Colony Stimulating
Factors including, but not limited to, Granulocyte Colony
Stimulating Factor (G-CSF), Macrophage Colony Stimulating Factor
(M-CSF or CSF-1), and GM-CSF; Epidermal Growth Factor (EGF) family
members including, but not limited to, EGF, HB-EGF, Amphiregulin,
Betacellulin, Epigen, Epiregulin, Neuregulin-3 (NRG-3), NRG1
isoform GGF2, NRG1 Isoform SMDF, NRG1-alpha/HRG1-alpha, NRG1-beta
1/HRG1-beta 1, TMEFF1, TMEFF2, and TGF-.alpha.; VEGF/PDGF family
members, including, but not limited to, Vascular Endothelial Growth
Factors (VEGF, otherwise known as Vascular Permeability Factor),
VEGF, VEGF-B, VEGF-C, VEGF-D; Placental derived Growth Factors
(PIGF), PIGF-1, PIGF-2, PIGF-3; Platelet-Derived Growth Factors
(PDGF), PDGF, PDGF-A, PDGF-B, PDGF-C, PDGF-AB; Neuropilin-1, and
Neuropilin-2; Transforming Growth Factor .beta. (TGF-.beta.) family
members, including, but not limited to, TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, TGF-.beta.4, and TGF-.beta.5; Schwann cell-derived
Growth Factor; Nerve Growth Factor (NGF); Insulin-like Growth
Factors 1 and 2 (IGF-1 and IGF-2); Glial Growth Factor; Tumor
Necrosis Factors TNF-.alpha. and TNF-.beta.; Connective tissue
growth factor (CTGF/CCN2); NOV/CCN3; PD-ECGF/gliostatin; Endocrine
Gland-derived Vascular Endothelial Growth Factor/prokineticin-1
(EG-VEGF/PK1); Hepassocin; Hepatocyte Growth Factor
(HGF/hepapoietin A/scatter factor); Beta subunit of Nerve Growth
Factor (.beta.-NGF); Progranulin; Thrombopoietin; Prolactin;
Prostaglandins; and Growth hormone (GH1 and GH2).
[0045] The method of the present invention uses known technologies
to immobilize enzymes, chelators, and antibodies in dialysis-like
cartridges has been developed (see, for example, Ambrus et al.
Science 201(4358): 837-839, 1978; Ambrus et al. Ann Intern Med
106(4): 531-537, 1987; Kalghatgi et al. Res Commun Chem Pathol
Pharmacol 27(3): 551-561, 1980) and is incorporated herein by
reference. An illustration of preparing proteins for immobilization
to the device developed, for example, by Aethlon Medical Inc. is
presented, for example, in U.S. Pat. Nos. 4,714,556 and 4,787,974,
5,528,057. Similar technologies can be used in the present
invention.
[0046] For example, for binding of affinity molecules to the
ultrafiltration membrane, matrix or other solid support, the
polymers of the solid support are first activated, i.e., made
susceptible for combining chemically with proteins, by using
processes known in the art. Any number of different polymers can be
used. To obtain a reactive polyacrylic acid polymer, for example,
carbodiimides can be used (Valuev et al., 1998, Biomaterials,
19:41-3.). Once the polymer has been activated, the affinity
molecules can be attached directly or via a linker. Suitable
linkers include, but are not limited to, avidin, streptavidin,
biotin, protein A, and protein G. For example, antibodies to
specific growth factors may be bound to streptavidin coated
polymers of the ultrafiltration membrane. The streptavidin coated
ultrafiltration membrane can also be used for the attachment of
oligonucleotide to which a biotin labeled base has been added to
the 3' end. The antibodies may also be directly bound to the
polymer of the ultrafiltration membrane using coupling agents such
as bifunctional reagents, or may be indirectly bound. For example,
Protein A or Protein G may be used to immobilize IgG against
specific growth factors.
[0047] For affinity absorbents, the solid support may be of various
shapes and chemical compositions. It may, for example, constitute a
column house filled with particulate polymers, the latter of
natural origin or artificially made. The particles may be
macroporous or their surface may be grafted, the latter in order to
enlarge the surface area. The particles may be spherical or
granulated and be based on polysaccharides, ceramic material,
glass, silica, plastic, or any combination of these or a like
material. A combination of these could, for example, be solid
particles coated with a suitable polymer of natural origin or
artificially made. Artificial membranes may also be used. These may
be flat sheet membranes made of cellulose, polyamide, polysulfone,
polypropylene or other types of material which are sufficiently
inert, biocompatible, non-toxic and to which the receptor could be
immobilized, either directly or after chemical modification of the
membrane surface. Capillary membranes, like the hollow fibers made
from cellulose, polypropylene or other materials suitable for this
type of membranes, may also be used.
[0048] In another embodiment the solid support is coated by ligands
which exhibit a specific interaction to the growth factor to be
removed from the blood circulation. Such ligands can be chosen from
a group comprising monoclonal antibodies including fragments or
engineered counterparts thereof, aptamers, peptides,
oligodeoxynucleosides including fragments thereof, intercalation
reagents including dyestuffs, oligosaccharides and chelating groups
interacting with the growth factor to be removed.
[0049] In another embodiment the adsorption device contains an
immobilized receptor binding specifically to the growth factor on
the solid support. Alternatively, the region of the receptor, that
is the peptide fragment, that is the site of binding for the growth
factor, may be used as an affinity compound. Any type of affinity
ligand/immobilized receptor combinations such as "antibodies and
antigens/haptens" and "protein and co-factors" could be used in
this application, provided that they exhibit a sufficiently high
binding affinity and selectivity to the growth factors and that the
ligand-receptor interaction is not interfered with by blood or
other body fluids or tissues being in contact with the adsorption
agent and/or the device.
[0050] In one embodiment, the affinity compounds bind to one or
more growth factors including, but not limited to, FGF-1/FGF
acidic, FGF-2/FGF basic, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7/KGF,
FGF-8, FGF-9, FGF-10, FGF-11, FGF-12, FGF-12, FGF-13, FGF-14,
FGF-15, FGF-16, FGF-17, FGF-18, FGF-19, FGF-20, FGF-21, FGF-22,
FGF-23, IL-1.alpha., IL-1.beta., IL-2, IL-3, IL-4, IL-5, IL-6,
IL-7, IL-8, IL-9, IL-10, IL-11, IL-12; G-CSF, M-CSF/CSF-1, GM-CSF,
EGF, HB-EGF, Amphiregulin, Betacellulin, Epigen, Epiregulin, NRG-3,
NRG1 isoform GGF2, NRG1 Isoform SMDF, NRG1-alpha/HRG1-alpha,
NRG1-beta 1/HRG1-beta 1, TMEFF1, TMEFF2, TGF-.alpha., VEGF, VEGF-B,
VEGF-C, VEGF-D, PIGF-1, PIGF-2, PIGF-3, PDGF, PDGF-A, PDGF-B,
PDGF-C, PDGF-AB, Neuropilin-1, Neuropilin-2, TGF-.beta.1,
TGF-.beta.2, TGF-.beta.3, TGF-.beta.4, TGF-.beta.5, Schwann
cell-derived Growth Factor, NGF, IGF-1 and IGF-2, Glial Growth
Factor, TNF-.alpha., TNF-.beta., CTGF/CCN2, NOVACCN3,
PD-ECGF/gliostatin, EG-VEGF/PK1, Hepassocin, HGF/hepapoietin
A/scatter factor, .beta.-NGF, Progranulin, Thrombopoietin,
Prolactin, Prostaglandins, GH1, GH2 and any combination thereof. In
a preferred embodiment, the affinity compounds bind to growth
factors comprising TGF-.alpha., TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2, FGF-1/FGF acidic, FGF-2/FGF
basic, VEGF, TNF-.alpha., FGF-7/KGF and any combination
thereof.
[0051] The anti-growth factor antibodies that can be used in the
methods of the present invention are commercially available from,
for example, R&D Systems (Minneapolis, Minn.), Abcam Limited
(Cambridge, UK), Sigma-Aldrich (St. Louis, Mo.) and Upstate
Biotechnology (Charlottesville, Va.). Alternatively, growth factor
antibodies may be produced by methods well known to those skilled
in the art. For example, monoclonal antibodies to growth factor
(preferably mammalian; more preferably human) can be produced by
generation of hybridomas in accordance with known methods.
Hybridomas formed in this manner are then screened using standard
methods, such as ELISA, to identify one or more hybridomas that
produce an antibody that specifically binds to a particular growth
factor. Full-length growth factors may be used as the immunogen,
or, alternatively, antigenic peptide fragments of growth factors
may be used.
[0052] As an alternative to preparing monoclonal antibody-secreting
hybridomas, a monoclonal antibody to a growth factor may be
identified and isolated by screening a recombinant combinatorial
immunoglobulin library (for example, an antibody phage display
library) to thereby isolate immunoglobulin library members that
bind to the growth factor. Kits for generating and screening phage
display libraries are commercially available from, for example,
Dyax Corp. (Cambridge, Mass.) and Maxim Biotech (South San
Francisco, Calif.). Additionally, examples of methods and reagents
particularly amenable for use in generating and screening antibody
display libraries can be found in the literature.
[0053] Polyclonal sera and antibodies may be produced by immunizing
a suitable subject, such as a rabbit, with a growth factor
(preferably mammalian; more preferably human) or an antigenic
fragment thereof. The antibody titer in the immunized subject may
be monitored over time by standard techniques, such as with ELISA,
using immobilized marker protein. If desired, the antibody
molecules directed the growth factor may be isolated from the
subject or culture media and further purified by well-known
techniques, such as protein A chromatography, to obtain an IgG
fraction.
[0054] Fragments of antibodies to a growth factor may be produced
by cleavage of the antibodies in accordance with methods well known
in the art. For example, immunologically active F(ab') and
F(ab').sub.2 fragments may be generated by treating the antibodies
with an enzyme such as pepsin. Additionally, chimeric, humanized,
and single-chain antibodies to a growth factor, comprising both
human and nonhuman portions, may be produced using standard
recombinant DNA techniques. Humanized antibodies to a growth factor
may also be produced using transgenic mice that are incapable of
expressing endogenous immunoglobulin heavy and light chain genes,
but which can express human heavy and light chain genes.
[0055] The subject treated by the methods of the present invention
may be a subject who is undergoing surgical wound healing. In one
embodiment, the surgery is for removal of a tumor. In one
embodiment, the tumor is a primary tumor. In one embodiment, the
subject is at risk for a tumor, such as a subject identified as
carrying tumor susceptibility mutations in genes such as BRCA1,
BRCA2, HPC1, MLH1, or MSH2. In one embodiment, the subject is at
risk for recurrence of cancer, for example, the subject was treated
for a cancer prior to infliction of the wound and is at risk for
recurrence of that cancer. In one embodiment, the surgery is
unrelated to tumor treatment. In one embodiment, the subject is
undergoing wound healing not induced by surgery, such as trauma
(for example, a broken bone, local burn, etc). For example, a
subject carrying a BRCA2 mutation is treated for injuries sustained
in a car accident concurrently with the methods of the present
invention.
[0056] Accordingly, the invention provides a method for reducing
the amount of at least one circulating growth factor from
circulating blood of a subject with a tissue injury, comprising
contacting at least a portion of the circulating blood of a subject
affected with a tissue injury with an extracorporeal adsorption
device wherein the device comprises at least one adsorption
compound that binds to at least one growth factor in the
circulating blood of the subject. In one embodiment, least one
growth factor is selected from the group consisting of TGF-.alpha.,
TGF-.beta.1, TGF-.beta.2, TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2,
FGF-1, FGF-2 (basic FGF), VEGF, TNF-.alpha., FGF-7 and any
combination thereof.
[0057] In one embodiment, the tissue injury is caused by surgery.
For example, the tissue injury can be induced during removal of a
tumor or caused by radiation therapy.
[0058] In one embodiment, the subject has or is at risk for
developing cancer.
[0059] The invention also provides a method of treating a subject
undergoing wound healing, comprising contacting the blood of said
subject with an extracorporeal adsorption device wherein the device
contains adsorption compounds that bind to growth factors in the
subject's blood. The subject is preferably affected with a surgical
wound. The surgical wound is preferably induced during surgical
removal of a tumor. In one embodiment, the subject with the tissue
injury is at risk for cancer. In one embodiment, the growth factors
are selected from the group consisting of FGF-1/FGF acidic,
FGF-2/FGF basic, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7/KGF, FGF-8,
FGF-9, FGF-10, FGF-11, FGF-12, FGF-12, FGF-13, FGF-14, FGF-15,
FGF-16, FGF-17, FGF-18, FGF-19, FGF-20, FGF-21, FGF-22, FGF-23,
IL-1.alpha., IL-1.beta., IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8,
IL-9, IL-10, IL-11, IL-12; G-CSF, M-CSF/CSF-1, GM-CSF, EGF, HB-EGF,
Amphiregulin, Betacellulin, Epigen, Epiregulin, NRG-3, NRG1 isoform
GGF2, NRG1 Isoform SMDF, NRG1-alpha/HRG1-alpha, NRG1-beta
1/HRG1-beta 1, TMEFF1, TMEFF2, TGF-.alpha., VEGF, VEGF-B, VEGF-C,
VEGF-D, PIGF-1, PIGF-2, PIGF-3, PDGF, PDGF-A, PDGF-B, PDGF-C,
PDGF-AB, Neuropilin-1, Neuropilin-2, TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, TGF-.beta.4, TGF-.beta.5, Schwann cell-derived Growth
Factor, NGF, IGF-1 and IGF-2, Glial Growth Factor, TNF-.alpha.,
TNF-.beta., CTGF/CCN2, NOV/CCN3, PD-ECGF/gliostatin, EG-VEGF/PK1,
Hepassocin, HGF/hepapoietin A/scatter factor, .beta.-NGF,
Progranulin, Thrombopoietin, Prolactin, Prostaglandins, GH1, GH2,
and any combination thereof. In one embodiment, the growth factor
is selected from the group consisting of TGF-.alpha., TGF-1.beta.,
TGF-.beta.2, TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2, FGF-1, FGF-2
(basic FGF), VEGF, TNF-.alpha., FGF-7 and any combination
thereof.
[0060] Alternatively, the growth factors are selected from the
group consisting of TGF-.alpha., TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2, FGF-1/FGF acidic, FGF-2/FGF
basic, VEGF, TNF-.alpha., FGF-7/KGF, and any combination
thereof.
[0061] The invention further provides a method of treating an
individual undergoing removal of a cancer comprising removing
undesired circulating growth factors from the individual. The
circulating growth factors are preferably removed by an
extracorporeal adsorption device. The device preferably comprises
at least one adsorption compound that binds to at least one growth
factor in the circulating blood of the individual.
[0062] In one preferred embodiment, the undesired circulating
growth factor is selected from the group consisting of TGF-.alpha.,
TGF-.beta.1, TGF-.beta.2, TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2,
FGF-1, FGF-2, VEGF, TNF-.alpha., FGF-7 and any combination thereof.
In an alternative embodiment, the growth factor is selected from
the group consisting of TGF-.alpha., TGF-.beta.1, TGF-.beta.2,
TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2, FGF-1, FGF-2 (basic FGF),
VEGF, TNF-.alpha., FGF-7 and any combination thereof.
[0063] The invention also provides for the use of an extracorporeal
device to remove at least one undesired circulating growth factor
from the blood of an individual with a tissue injury, such as
tissue injury icaused by surgery or radiation therapy. In one
preferred embodiment, the subject is at risk for cancer. In another
embodiment, the subject has or has had a cancer, or a benign tumor.
In one preferred embodiment, the undesired circulating growth
factor is selected from the group consisting of TGF-.alpha.,
TGF-.beta.1, TGF-.beta.2, TGF-.beta.3, PDGF, EGF, IGF-1, IGF-2,
FGF-1, FGF-2, VEGF including at least one VEGF sub type,
TNF-.alpha., FGF-7 and any combination thereof.
[0064] In conjunction of the methods of the present invention, the
subject may also receive additional treatment modalities or
therapeutics for the treatment of cancer or other condition. For
example, the subject may undergo treatment for cancer including the
taking of cytotoxic agents or chemotherapeutic agents
simultaneously or concurrently with the methods of the present
invention.
[0065] Cytotoxic agents include any substance that inhibits or
prevents the function of cells and/or causes destruction of cells.
Cytotoxic agents include radioactive isotopes, chemotherapeutic
agents, and toxins such as, but not limited to, active toxins of
bacterial, fungal, plant or animal origin, or fragments thereof.
Some radionuclides, like indium-111, are used as diagnostic agents
and are as such administered with low activity, but could also be
used for therapeutic purposes if given in higher doses and are
therefore also referred to as cytotoxic agents herein.
[0066] Chemotherapeutic agents are chemical compounds useful in the
treatment of cancer. Examples of chemotherapeutic agents include
Adriamycin, Doxorubicin, 5-Fluoruracil, Cytosine arabinoside
("Ara-C"), Cyclophosphamide, Thioptepa, Busulfan, Cytoxin, Taxol,
Methotrexate, Cisplatin, Melphalan, Vinblastine, Bleomycin,
Etoposide, Ifosfamide, Mitomycin C, Mitoxantrone, Vincristine,
Vinorelbine, Carboplatin, Tenisposide, Duanomysin, Carminomycin,
Aminopterin, Dactinomycin, Mitomycins, Esperamicins (see U.S. Pat.
No. 4,675,187), Maytansinoids, Melphalan and other related nitrogen
mustards.
[0067] In one embodiment, reduction of growth factors from systemic
circulation means a reduction of at least about 5-10%, 10-25%, and
in increasing preference, reductions of at least about 50%, 60%,
70%, 80%, 90% and 95%. Levels of circulating growth factor may be
evaluated using any known technique, such as enzyme-linked
immunosorbent assays (ELISA) or radioimmunoassays (RIA).
[0068] For the method of the present invention, blood is withdrawn
from a subject and contacted with the extracorporeal adsorption
device. Blood access may be achieved through peripheral vein
catheters or, if higher blood flow is needed, through central vein
catheters such as, but not limited to, subclavian or femoral
catheters. The adsorption device can be directly perfused with
blood from subjects and returned to the subjects without further
manipulations. Alternatively, blood can be separated into plasma
and cellular components by standard techniques. The plasma is then
contacted with the adsorbent compounds to remove the growth factors
by binding between growth factor and adsorbent compound. The plasma
can then be recombined with the cellular components and returned to
the subject. Alternatively, the cellular components may be returned
to the subject separately. In one embodiment, at least 1,
preferably 2, more preferably 3, 4, 5, 6 or even more volumes of
blood are passed through the extracorporeal adsorption device. The
treatment can be repeated periodically until a desired response has
been achieved. For example, the treatment can be carried out for 4
hours once a week. Growth factor levels can be assessed in the
effluent from the adsorption device by standard techniques such as
ELISA and RIA.
[0069] All patents, patent applications and publications cited
throughout the specification are incorporated herein by reference
in their entirety.
* * * * *